Estrogenic compounds and animal growth promoters



United States Patent Ofifice 3,373,033 ESTROGENIC COMPOUNDS AND ANIMALGROWTH PROMOTERS' Edward B. Hodge, Phil H. Hidy, and Herbert L.Wehrmeister, Terre Hante, Ind., assignors to Commercial SolventsCorporation, a corporation of Maryland No Drawing. Continuation-impartof application Ser. No. 561,373, June 29, 1966. This application Mar. 3,1967, Ser. No. 620,270.

10 Claims. (Cl. 99-2) ABSTRACT OF THE DISCLOSURE where A is selectedfrom the group consisting of --CH=CH- and CH CH R is selected from thegroup consisting of hydrogen, lower alkyl, benzyl and lower saturatedacyclic acyl; X is selected from the group consisting of hydrogen, SO-H, -SO R --SO Cl and -SO NHR wherein R is lower alkyl and R is selectedfrom the group consisting of hydrogen, alkyl of about 1 to 20 carbonatoms, monocyclic aryl and halogen with the proviso that not more thanone X is hydrogen and that R is selected from the group consisting oflower alkyl and benzyl when X is --SO Cl, and B is selected from thegroup consisting of C=O, CHOH and CH with the proviso that A is CH --CHwhen B is CH and animal feeds containing growth promoting amountsthereof.

where A is --CH CH or CH=CH-; R is hydrogen; substituted orunsubstituted alkyl, e.g. lower alkyl such as methyl, ethyl, hexyl,etc.; acyl, e.g., lower saturated acyclic acyl radicals such as acetyl;or aralyl, e.g., benzyl .and bromo benzyl; X is hydrogen, SO H, SO Cl,SO R or -SO NHR where R is alkyl, e.g., lower alkyl of from about 1 to 8carbon atoms, and R is hydrogen, halo, such as chloro, a saturatedaliphatic radical of 1 to 20 carbon atoms, the substituents of which donot interfere with the sulfonation reaction, especially lower alkyls,aryl such as monocyclic aryls including phenyl and substituted phenyl,the substitutents of which do not interfere with reaction such aschlorophenyl and nitrophenyl; with the provisos that not more than one Xis hydrogen and that when X is SO Cl, R is lower alkyl 3,373,033Patented Mar. 12, 1968 or benzyl; and B is C=O, CHOH or CH with theproviso that when B is CH A is -CH -CH The compounds can be administeredto animals by any suitable method including oral and parenteraladministrations. For example, the compound can be blended with ordinaryfeed containing nutritional values in an amount sufii'cient to producethe desired rate of growth and can thus be fed directly to the animals,or the compound can be suspended in a suitable injection suspensionmedium such as peanut oil and injected parenterally. The amount ofcompound fed to an animal, of course, varies depending upon the animal,desired rate of growth and the like.

When the new compounds are to be administered in feeds, an animal feedcomposition may be prepared containing the usual nutritionally-balancedquantities of carbohydrates, proteins, vitamins and minerals, togetherwith the compound of the present invention. Some of these usual dietaryelements are grains, such as ground grain and grain lay-products; animalprotein substances, such as those found in fish meal and meat scraps,vegetable proteins like soybean oil meal or peanut oil meal;vitaminaceous materials, e.g. vitamin A and D mixtures; riboflavinsupplements and other vitamin B complex members; and bone meal andlimestone to provide minerals. A type of conventional feed material foruse with cattle includes alfalfa hay and ground corn cobs together withsupplementary vitaminaceous substances if desired.

The compounds of the present invention are produced by sulfonation ofthe compound:

hereinafter referred to as the fermentation estrogenic sub stance(F.E.S.) to add one, or more, sulfonic acid radicals to the benzenering. F.E.S. sulfonamides can be produced by reaction of the PBS.sulfonic acid chloride with an amine, i.e., of the formula NH R where Rhas the above meaning. The alkyl esters of F.E.S. sulfonic acids, whereX is SO R are produced by reaction of the F.E.S. sulfonic acid chloridewith an alkanol of the formula R OH where R has the above meaning or byheating the sodium salt of the sulfonic acid with an alkyl sulfate. TheF.E.S. sulfonic acids of this invention are also useful as intermediatesin the production of nitro or amine substituted F.E.S. compounds asshown in US. patent application Ser. No. 561,369, filed on even dateherewith.

It is desirable to protect the hydroxyl groups of F.E.S. againstundesirable side reactions before sulfonation which can be accomplishedby converting the hydroxyl groups to ethers, e.g., methyl ethers, astaught in US. Patent 3,239,- 342 and subsequently removing the maskinggroup to regenerate the hydroxyl group. The hydroxyl group can beregenerated from, for example, a methyl ether by hydrolysis or byrefluxing in acetic acid solution with hydrobromic acid. Aluminumchloride is another effective dealkylating agent. Other suitable maskinggroups include generally lower alkyls and benzyl. Compounds where R isbenzyl are described in copending application Ser. No. 532,113, filedMar. 7, 1966, now abandoned, and application Ser. No. 620,259, filedMar. 3, 1967, US. Patents 3,239,354; 3,239,347 and 3,239,356, forexample, describe dihydro F.E.S. compounds where A is CH CH and R isacyl or a combination of alkyl and acyl. Tetraether of F.E.S.,hydrolyzing the ether and subsequently acylating the compound.

The following examples are offered to illustrate this invention;however, the invention is not limited to the specific materials,amounts, and procedures set forth.

Example I Five milliliters of 100% H 80 (equal volumes of 97% H 80 and20% oleum) were cooled in an ice salt bath until it solidified and then1 g. of F.E.S. was added to the frozen mass and mixed to form a mush.The temperature was allowed to rise until the acid melted and thereaction mixture was mixed until all F.E.S. had dissolved. Afterstanding 2 hours at refrigerator temperature, the solution was pouredonto ice to yield a precipitate which was separated by decantation.

The precipitate was dissolved in sufficient water to form a.non-gelatinous solution and treated with char to decolorize. Char wasremoved by filtration and the product was recrystallized by addition ofconcentrated HCl until about one third volume of HCl had been added pervolume of solution. When crystallization was complete, the product wascollected by filtration and dried in vacuo to yield 492 mg., M.P.114-115 C.

Analysis C18H2203S.1%H2P Calculated Found Percent O 50.81 50.05 PercentH 5.92 6.14 Percent O 35. 72 35. 82 Percent S 7. 53 7. 68

Example II Dihydro F.E.S., tetrahydro F.E.S., deoxy tetrahydro F.'E.S.,F.E.S. diethyl ether, dihydro F.E.S. dimethyl ether, and F.E.S. 4-benzylether are sulphonated by the process of Example I at a somewhat highertemperature.

Example III F.E.S.. 2,4-dimethyl ether sulfonic acid is producedaccording to Example I, substituting F.E.S. 2,4-dimethyl ether forF.E.S., and then reacted in aqueous solution with sodium hydroxide toproduce the sodium salt of F.E.S. 2,4-dimethyl ether sulfonic acid. Areaction mixture of 3 mols F.E.S. dimethyl ether sulfonic acid sodiumsalt and 1 mol PCl is heated at 170-180 C., to produce the correspondingacid chloride. The reaction mixture is. cooled and the acid chlorideseparated and dried.

F.E.S. sulfonic acid chlorides are also produced according to theprocess of Example III substituting dihydro F.E.S. 2,4-dimethyl ether,tetrahydro F.E.S. trimethyl ether and deoxy F.E.S. 2,4-dimethyl ether.

Example IV F.E.S. 2,4-dimethyl ether sulfonic acid chloride is stirredwith methanol to produce the methyl ester of F.E.S. 2,4-dimethyl ethersulfonic acid, which is recovered.

Example V F.E.S. 2,4-dimethyl ether sulfonic acid chloride is shakenwith aqueous ammonia to'produce F.E.S. 2,4-dimethyl ether sulfonamidewhich crystallizes from alcohol and has the formula 4.- Example VI Thefollowing amines are substituted for ammonia in the process of Example Vto produce the corresponding F.E.S. 2,4-dimethyl ether sulfonamide:methylamine, ethylamine, n-amylamine, aniline, chloroaniline,nitroaniline and laurylamine.

Example VII F.E.S. 2,4-dimethyl ether sulfonamide is reacted with sodiumhypochlorite to produce the chloroamide Example VIII Dihydro F.E.S.2,4-dimethyl ether sulfonic acid chloride is substituted for F.E.S.2,4-dirnethyl ether sulfonic acid chloride in the procedure set forth inExample V to produce a compound having the formula (H) OH;

CHs-O The corresponding compound is produced from deoxy F.E.S.

Example IX The sulfonation porduct of Example V is demethylated byheating at 120 C. in benzene solution with two equivalents of aluminumchloride and the demethylated prod uct is acylated with acetic anhydrideto produce the corresponding 2,4'diacetyl substituted F.E.S.sulfonamide.

Example X Six head of cattle are fed a daily ration including a mixtureof alfalfa hay and ground corn cobs containing from 1 to 20 ounces ofthe compound produced in Example I per hundred pounds of ration andtheir rate of growth is increased.

Example XI For young beef cattle, i.e., calves to yearlings running totwo year olds, each animal is given 5 to 20 milligrams per day of thecompound produced in Example I intimately admixed in about 18 to 22pounds per head per day of a complete pelleted'ration for about ldays.The complete pelleted ration includes in addition to the compound ofExample I the following:

from 5 to 90 milligrams per head per day.

Grams/ton: Mg./head/day .5 5 1.0 2.0 20 4.0 40 8.0 80

These gram amounts are premixed with, for example, 10 pounds of soybeanhulls prior to admixture with the other ingredients.

Example XIII For young swine, i.e., six week old pigs to about 100 poundpigs, each animal is given 5 to 20 milligrams per day of the compound ofExample IV intimately admixed in about 1 /2 to 5 /2 pounds per head perday of a grower ration until it reaches a weight of about 100 pounds.When the swine weigh between 90 and 125 pounds the feed is changed toone whereby each amimal is given 20 to 50 milligrams per day of thecompound of Example IV intimately admixed in about 5 /2 to 10 pounds perhead per day of a finisher ration until it reaches market weight ofabout 220 pounds. The grower and finisher ration include in addition tothe compound of Example IV the following:

Grower, Finisher,

Percent Percent Ground Yellow Corn 77 86. 7 Soybean Meal (44% protein)16 6. 5 Meat and Bone Scraps (50% protein) 2. 5 2. 5 Dehydl'ate AlfalfaMeal (17%) 2. 5 2. 5 Steamed Bone meal 0. 5 0. 5 Ground Limestone 0.5 0.3 Iodized Salt 0. 5 0. 5

Vitamin, Antibiotic and Trace Mineral Premix 0.5 0. 5

The compound of Example IV is admixed with the above ingredients in ablender in the following amounts in milligrams per pound to provide anappropriate feed with dosage levels ranging from 6 to 96 milligrams perhead per day.

Mg./pound: Mg./head/day 2 6 4 12 8 24 16 48 32 96 Example XIV For 4 to10 month old lambs weighing 50 to 70 pounds, each animal is given 1 tomilligrams per day of the compound of Example V, intimately admixed inabout 3 to 6 pounds per head per day of a complete ration for 30 to 90days. The complete ration includes in addition to the compound ofExample V the following:

The compound of Example V is admixed with the above ingredients in ablender in the following amounts in milligrams per pound to provide an.appropriate complete feed with dosage levels ranging from 1 to 15milligrams per head per day.

Mg./pound Mg./head/day Example XV For broilers, i.e., day old to fourweek old chicks, a

grower feed is prepared for feeding to the chicks for the first fourweeks and a finisher feed is prepared for feeding the four week oldchicks for the last five weeks until they reach market weight of two anda half to three pounds dressed. For each pound of weight gain, thechicks eat about 1.5 to 2 pounds of feed. Thus they eat about 1.5 poundsof feed during the first four weeks and about five pounds of feed duringthe next five weeks. During the course of this feeding schedule, eachbird should receive a total of about 12 to 36 milligrams of the compoundof Example VII in the grower and finisher feed each of which includes inaddition to the compound of Example VII the following:

Grower Finisher (lbs) (lbs) Ground Yellow Corn 1, 00!) 1, 200 Soybeanmeal (44% Protein) 700 500 Fish Meal (60% Protein) 100 Alfalfa Meal 5050 Meat and Bone Scraps 0 80 Animal Fat 80 80 Dicalcium Phosphate. 35 40Iodized Salt 10 10 Limestone 15 Premix Vitamins Trace Minerals andAntioties 10 1 0 where A is selected from the group consisting of -OH-*CH- and --CH CH R is selected from the group consisting of hydrogen,lower alkyl, benzyl and lower saturated acyclic acyl; X is selected fromthe group consisting of hydrogen, 'SO H, -SO R SO Cl and -SO NHR whereinR is lower alkyl and R is selected from the group consisting ofhydrogen, alkyl of about 1 to 20 carbon atoms, monocyclic aryl andhalogen with the provisos that not more than one X is hydrogen and thatR is selected from the group consisting of lower alkyl and benzyl when Xis S0 01, and B is selected from the group consisting of C=O, CHOH andCH with the proviso that A is CH -CH when B is CH 2. The compound ofclaim 1 wherein A is OH=CI-I, B is C=O R and Y are hydrogen, and X is-SO H.

3. The compound of claim 1 wherein A is -CH=CH, B is C=O R and X arehydrogen, and Y is -SO H.

4. The compound of claim 1 wherein 'SO H and Y is hydrogen.

7 8 S. The compound of claim 1 wherein X is -SO R and growth promotingamounts of the compound of 6. The compound of claim 1 wherein X is SOCl. claim 2. 7. The compound of claim 1 wherein X is S0 NHR Noreferences cited.

8. The compound of claim '1 wherein X is SO H. 9. An animal feedcomprising anutritional diluent and 5 LOUIS MONACELL, PrimaryExaminergrowth promoting amounts of the compound of claim 1. KLARE 1HExamine .10. An animal feed comprising a nutritional diluent UNITEDSTATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. "5,373,033March 12 1968 Edward B. Hodge et 211.

It is certified that error appears in the above identified patent andthat said Letters Patent are hereby corrected as shown below:

Column 1, line 60, "aralyl" should read aralkyl line 68, "substitutents"Should read substituents Column 3, line 26, in the table, "H P" shouldread H O Column 4, line 34, "porduct" should read product Column 5,1ine.27, "amimal" should read animal line 69 "cronn should read cornColumn 6 line 74, after "wherein" insert X is Column 7, line 4, "SO H"should read SO H Signed and sealed this 7th day of October 1969.

(SEAL) Attest:

Edward M. Fletcher, Jr. E.

Attesting Officer Commissioner of Patents

